Method for killing fungi with betaamino-ethyl ketones



United States Patent O M 3,257,274 I METHOD FOR KILLING FUNGI WITH BETA-AMINO-ETHYL KETONES Nicola Loprieno, Pisa, Emilio Plastino, Florence,and

This invention relates to beta-aminoethylaryl ket'one fungicides havinga presentive activity by contact and by fumigation and/or anendophytotherapeutic and immunizing activity.

The protection of agrarian cultures a gainst pathogenic agents belongingto the vegetal kingdom (fungi, bacteria) has heretofore been effectedmainly by means of parasiticidal chemical compounds exerting apreventive surface or covering action. In addition to the classicalfungicides on the basis of copper, sulfur and mercury, fungicides on thebasis of new'o'rganic compounds have recently been utilized. As isknown, dithiocanbamates have assumed a prominent position among thoseorganic fungicides. This was due to their high activity, easyapplication, absence of phytotoxicity and above all because they madethe production of fungicides economical by rendering it independent ofthe uncertain copper supply. It is also known that 'while theintroduction of the new acupric fungicides represented considerableprogress in the field of phytoiatry, several technical and economicalproblems are still open.

With covering fungicides having preventive contact action, farmers haveto keep the plants constantly covered with a film of fungicidal activeingredient, starting the treatments before infestation occurs andrenewing, by frequent applications, the protective layer throughout theentire period during which infections can occur; this period almostalways coincides with the entire vegetative stage.

Disregardin-g the effect of atmospheric factors on the degradation andremoval of the active ingredient, the repetition of the treatments isparticularly necessary in the springtime 'when plants exert an intensegrowth activity, in order to protect the plants as they grow. When wealso consider that in some stages of the vegetative cycle of theagrarian culture, rainfall, continuous or intermittent, prohibits thecovering treatments while at the same time being extremely favorable forthe propagation of the infections, it becomes obvious that the methodsand means having a preventive external action, and also thephytosanitary problems to which phytoiatric science and technique havemade contributions, cannot be considered solved in a technically andeconomically satisfactory manner.

To complete the picture, it should be added that the aforementionedmethods and means, the only ones available to the farmer for protectingcultures, do not make it possible to control some of the most seriousparasitic forms, which not only damage the crops but also inevitablylead to the destruction of the plants themselves. These diseases, due totheir vascular development, are called systemic diseases by someauthors. The numerous tracheomycoses caused by various fungus species,such as Fusarium, Ver'ticellium, Deuterophoma, Graphium must bementioned in particular.

At this point, it is evident that a substantial improvement in thefungicidal means and methods now used and solution of questions involvedin the control of systemic diseases can be obtained only if chemicalsubstances penetrating the vegetal organism in various ways areavailable. These chemical substances either by acting locally or farfrom the application point must be capable of destroying the alreadypresent parasite or of preventing its development in the inner parts ofthe plant.

The s-o-called systemic insecticides were already used in the pestcontrol, thus demonstrating theposs-ibility of introducing chemicalsubstances in various ways inside the vegetal organism, with therelative displacement and diffusion of the substances in the variousorgans and tissues 'of the plant. A

While the control of animal parasites (insects, mites, etc.) inside theplants by means of systemic substances is obviously facilitated by thesubstantial differences existing between the physiological andbiochemical processes of the parasites and the host plant, thisdistinction almost vanishes in case of endotherapeutic fungicides orbactericides. y

In fact, it is obvious that the anichlolinesterasic activity to whichthe lethal action of some systemic insecticidal and miticidal phosphoricesters is essentially to be ascribed, does not interfere with the vitalprocesses of the higher plants and said substances do not exert anyphy'totoxic activity.

Endotherapic fungicides or bacter'icide's, on the contrary, can easilyhave a negative effect on the treated plants since the higher vegetalorganisms (agrarian plants) must be suspected of a higher physiologicaland biochemical afiinity for the fungicides and bactericides than theorganisms belonging to the lowest order of the vegetal kingdom (fungi orbacteria).

An object of this invention is the utilization of El-aminoethyl'arylketones as endophytotherapeutic agent's.

Another object of this invention is the endophytotherapy of fungusdiseases by means of products that do not have phytotoxic action.

It should be stated in advance that we consider a chemical product asend'otherapeutic and immunizing when the (product, without causingappreciable phytotoxic action, is capable of penetrating through theouter protective plant layers (cuticle, epidermis, periderm), diffusinginto the inner tissues and of being transferred more or less rapidlyfrom one organ 'to another of the plant. The product destroys, both atthe point of application and at locations removed therefrom, thernycelium of the already installed pathogen (curative action) or thepromycelic tube, coming from spores germinated outside, as soon as it isintroduced into the host (immunizing action).

The same products have a consider-able preventive covering andfumigating activity against the fungus spores before the latter causeinfection.

We have found that the fi-aminoethylarylketones are products of thehighest interest from the aforeconsidered points of view.

The compounds of the aforementioned groups are comprised in the generalformula in which A is an aryl that can be variably substituted, and Y isan amine or an amine salt. The salts (e.g., picrates, ferrocyanides) arepractically water insoluble and are particularly useful as surfaceprotectant fungicides because they have a very high immediateeffectiveness and a satisfactory resistance to removal by rain.

In Table I are listed the compounds of the aforementioned class whichare of particular interest, together with their physico-chemicalcharacteristics, the mark with which they will be indicated hereinbelow,and representative formulas.

The compounds listed in Table I were s'ynthetized according to theMannish reaction, i.e., by reacting an arylmethylketone (ACO-CH in whichA has the afore- 3 mentioned meaning) with an amine salt andformaldehyde. (Mannich C. B'er., 55 (1922), 356, 3510.)

This method has been improved by other authors (Blicke, F. F.,Burkhalter, J. H., J. Am. Chem. Soc, 64 (1942), 453. Fry, E. M., J. Org.Chem., (1945), 259. Maxwell, C. E., Organic Syntheses, 1946, vol. 23,page 30. Winstein, S., J. Org. Chem, 11 (1946), 215).

The corresponding free bases are obtained by known methods, namely byremoval of hydrochloric acid with sodium hydroxide from an aqueoussolution of the hydrochloride and solvent extraction.

The practically water insoluble salts (e.g. picrates, ferrocyanides) canbe obtained by precipitation with the corresponding acids from aqueoussolutions of the hydrochloride salts.

TABLE I.5-AMINOETHYLARYLKETONES OF THE FORMULA A-CO-CH2CHz-Y MarkCompound Physical characteristics Analytical data,

percent S. fi-dimcthylaminoethylphenyl ketone hydrochloride Crystals,M.P. 155156 C N=6.59 (calc. 6.55).

O Cl=16.69 (calc. 16.69). H Qo-omommcrrmnor S. 92.B-dimethylaminoethyl-2-hydroxyphcnyl ketone hydrochloride Crystals, M.P.173174 C N=6. (6.12).

Ol=l5.62(l5.43).

S. 113 fl-isopropylaminoethylphenyl ketoue hydrochloride Crystals, M.P.175 C N=6.30(6.15).

Cl=63.33 (63.29) H=8.02(7.95)

S. 114 B-piperidinoethylphenyl ketone hydrochloride Crystals, M.P.192-193 C N=5.71(5.52).

H01 O CH2CH2 ll CCH2CH2N\ CH2 CHr-Cz (B-piperidylethylphenyl kctonehydrochloride).

S. 116 B-piperidinoethylphenyl ketone (B-piperidylcthylphenyl ketone)0il. N=6.52(6.44).

S. 117 B-morpholinoethylphenyl ketone hydrochloride Crystals, M.P.ISO-181 C N=5.71'(5.74).

3101 (H) /CH2CEZ C (JCH2CHzN\ GET-CH2 S. 123B-piperidinoethyll-chlorophcnyl kctone (fi-piperidylethyl-4-chlorophenylCrystals, M.P. 194194.5 C N =5.11(4.86). Cl=24.0(24.6).

8.127 B-isopropylaminoethyl-4-chlorophenyl ketonc hydrochlorideCrystals, M.P. 167168 C N=4.9s(5.34).

. fl-isopropylaminoethyl3,4-dichlorophenyl kctonc hydrochlorideCrystals, M.P. 193-194 C 137 5-dimethylaminocthyl-4-ohlorophenyl kctonchydrochloride Crystals, M.P. l71173 C N =5.70(5.64)

S. 138 B-dimethylaminccthyl-3,4-dioh10r0phenylkctonc hydrochlorideCrystals, M.P. HIS-195 C N =5.02(4,95).

S. 139 fl-morpholinoethyl4-chlor0phenyl ketonc hydrochloride Crystals,M.P. 205-2116 C C1=24,46 (24.78),

S. 142 fl-dimethylaminocthyl-4-mcthylphenyl kctone hydrochlorideCrystals, M.P. 156156.5 C N=6.14(6.15).

S. 143 B-dimethylaminoethyl-3,4-dichlorophcnyl kctone fcrrocyanideCrystals, M.P. 158 C i [Cl 3-C-CH2CHN(CHs)2]a-H3[Fc(CN)s] S. 145B-dimethylaminoethyl-fa,4-dichl0rophcnyl ketone oxalate Crystals, M.P.loll- C N=4.45(4.16).

fl) [OlCCHzCHz-N(CHa)z]z-(COOH) S. 147 fl-dimcthylaminoethyl-2-thienylketone hydrochloride Crystals, M.P. 182-183 C N=6.65(6.37)

. S=l4.90(l4.59) CS\ C Cl=16.5 3(16.13) H CCCHzCHzN(CH -HCl C C S. 149[El-morpholinoethyl-3,4-dichl0rophcnyl kctonc hydrochloride Crystals,M.P. -181 C N=4,50(4 37) Ol=3l.20(33.18).

TABLE I-Continued Mark Compound Physical characteristics Analyticaldata,

percent S. 161. fl-dimcthylaminocthyl-3-nitrophenylketone hydrochloride;Crystals, M.P.197-198 C N=11.07(l0.82). Cl=13.60(13.70).

S. 169 fi-dimethylaminoethyl 3nitro-4-chlorophenyl ketone hydrochlorideCrystals, M.P. 178 C 1g1=9 .;7-51(6?(.55

C=45.16(45.0 V H=4.83(4.8l).

S. 174 B-diethylaminoetliyl-3A-dichlorophenyl ketone hydrochlorideCrystals, M.P. 130-132 C N=4.64(4.51).

S. 178 fl-dimethylaminoethyl-3-nitro-4-methylphenylketone hydrochlorideCrystals, M.P. 176-177 C N =10.39(10.27).

S. 179 fl-isopropylaminoethyl-3-nitrophenyl kctone hydrochlorideCrystals, M.P. 163-165 C N =10.22(10.20).

S. 180 fi-diethylamin0ethy1-3-nitrophenyl ketone Crystals, M.P. 136-138C N=9.90(9.72) S. 18119-15opropylaminoethy1-3-nitl0-4-chlorophcnylketone hydrochlorideCrystals, M.P. 176-178 0.. N=9.16(9.11).

S. 185 fl-diethylaminoethyl--chlorophenyl ketone hydrochloride Crystals,M.P. 137-138 C N=5.23(5.11).

s. 187. fl-isopropylaminoethyl-4-methylpheny1kotone hydrochlorideCrystals, M.P. 171-172 C N =6.0(5.79).

S. 192 fl-dimethylaminocthyl-2,4-dichlorophenyl ketone hydrochlorideCrystals, M.P. 138-139" C N=5.08(4.96).

S. 196 fl-isopropylaminoethyl-3-nitr0-4-methylphenyl ketonehydrochloride Crystals, M.P. 171-172" C N=9.92(9.80).

S. 201 fl-dimethylaminoethy1-2-(tctrahydronaphthyl) ketone hydrochlorideCrystals, M.P. 160-161 C N =5.49(5.23).

CHC-CH2-CHzN(CH -HCl CH2 J CH2 S. 202B-pr0pylaminoethyll-chlorophenylketone hydrochloride Crystals,M.P.140-142" C N=4.87(4.60).

Cl=ll.60(11.65).

s, 205 B-dimethylaminoethyl-4-bromo-1-naphthyl ketone hydrochlorideCrystals, M.P. 176-177 C l 4 CH3 Br=23.44(23.32). (I) OCH2CH2N v CH HCl

I Br

S. 209 fi-dimethylaminoethyl-B-bromophcnyl kctone hydrochlorideCrystals, M.P. 205-206 C N=5.l0(4.78).

. Cl=11.98(l2.11). S. 210 fi-dimethylaminoethyl-l-naphthyl ketonehydrochloride Crystals, M.P. ISA-155 C N=5.58(5.31).

. Ol=13.52(13.44). S. 211 fl-dimethylaminoethyl-2-naphthyl ketonehydrochloride Crystals, M.P. 170171 C N=5.39(5.33).

S. 212 fl-dimethylamin0ethyl-4-chloro-l-naphthyl ketone hydrochlorideCrystals, M.P. 154-155 C N=4.88(4.69).

S. 213 fl-dimethylaminoethyM-methoxyphenyl ketone hydrochlorideCrystals, M.P. 183-184" C N=6.1(5.74).

S. 214 fl-dimethylaminoethyl-4-nitro-1-naphthyl ketonc hydrochlorideCrystals, M.P. 200-202" C lll=8. COCHz-CHZN(CH3)2-HCI S. 215fi-dipropylaminoethyl-l-naphthyl ketone hydrochloride. Crystals, M.P.127-128 C N=4. 42(4. 38).

, Cl=1l. 13(11. 08

S. 216 B-isopropylaminoethyl-l-naphthyl kctone hydrochloride Crystals,M.P. 156-158 C N=5. 36(5. 04).

S. 217 B-dibutylaminoethyl-l-naphthyl ketone hydrochloride Crystals,M.P. 100-101 C N =4. 42(4. 38).

S. 218 fl-diethylaminoethyl-l-naphthyl ketone hydrochloride Crystals,M.P. 134-l35 C N=5. 2(4. 80).

Cl=l2.36(12.15).

S. 219 fi-morpholinoethyl-1-naphthyl ketone hydrochloride Crystals, M.P.171-172 C N =4. 76(4. 58). 01-11. 86(11, 5 S. 220.00fl-piperidine-ethyld-naphthyl ketone hydrochloride (fl-piperidylethyl-l-Crystals, M.P. 176177 C N =4. 94(4. 61).

naphthyl ketone hydrochloride). Cl=l1. 62(11. 67).

S. 232 B-piperidinoethyl-2-hydroxyphenyl ketone hydrochloride(B-piperidyl- Crystals, M.P. 170-171 C N=5.51(5.19).

ethyl-2-hydroxyphenyl kctohe hydrochloride).

TABLE IContinucd Mark Compound Physical characteristics Analytical data,

. percent S. 233 B-diethylaminocthyl-4-chloro-1-naphthy1 ketonehydrochloride Crystals, M.P. 126127 C N=4.60(4.29)

S. 234 fl-dipropylaminoethyl-4-chloro-1-naphthyl ketone hydrochlorideCrystals, M.P. 111-113 C N=4.28(3.95).

S. 236 fl-morpholinoethyl-4-chloro-1-naphthyl ketone hydrochlorideCrystals, M.P. 176-176.5 C N=4.37(4.11).

S. 237- fl-piperidinoethyl-4-chloro-1-naphthyl ketone hydrochloride(B-piperidyl- Crystals, M.P. 176-177 C N =4.48(4.31).

ethyl-4-chloro-1-naphthyl ketone hydrochloride). Cl=21.05(20.96).

S. 239. B-morpholinoethyl-2,4-dich1orophenyl kctone hydrochlorideCrystals, M.P. 196-197" C N =4.48(4.31).

S. 240 fi-piperidinoethyl-2,4-dichlorophenyl ketone hydrochloride(fl-piperidyl- Crystals, M.P. 163-164 C N=4.66(4.37).

ethyl-2,4-dichlorophenyl ketone hydrochloride).

S. 251 fi-diethylaminoethyl-2,4-dichlorophenyl ketone hydrochlorideCrystals, M.P. 149-150 C N=4.83(4.48).

S. 252 fl-piperidinoethyl-S-nitrophenyl kctone hydrochloride(fi-piperidylethyl-Zi- Crystals, M.P. 173-175 C N'=8.95(9.37).

nitrophenyl ketone hydrochloride).

S. 253 B-morpholinoethyl-IS-nitrophenyl ketone hydrochloride Crystals,M.P. 185186 C N=9. 57(9. 31).

S. 255 fi-morpholinoethylA-methylphenyl ketone hydrochloride Crystals,M.P. 205 C N=5. 41(5. 19).

S. 256 B-piperidinoethyll-methylphenyl ketone hydrochloride(fl-piperidylethyl- Crystals, M.P. 173-174 C N =5. 40(5.23).

4-methylphenyl ketono hydrochloride).

S. 259 B-dimcthylaminoethyl-l-naphthyl ketone oxalate Crystals, M.P.137-140 C S. 261 B-piperidinoethyl-3,4-dichlorophenyl ketonchydrochloride (fi-p poridyl- Crystals, M.P. 175-176 C Cl=32,90(32.97).

ethyl-3,4-dichloropheny1 ketone hydrochloride).

S. 262 B-morpholinoethyl-3-nitro-4-ch1orophenyl ketone hydrochlorideCrtystals, M.P. 168-169 C Cl=21.03(21.15).

S. 263 fl-piperidinoethyl-3-nitro-4-chlorophenyl ketone hydrochloride(fl-p pcri- Crystals, M.P. l79-180 C Cl=20.95(21.28).

dyletliyl-3-nitro-4-chlorophenyl kctone hydrochloride).

S. 272 B-dirnethylaminoethyl-I-naphthyl ketone Oil S. 279B-piperidinocthyl-4-ch1oro-1-naphthyl ketone(fl-piperidylethyl-4-chloro-l- Oil naphthyl kctone).

S. 280 fi-dimethylaminoethyl-Q-anthranyl ketone hydrochloride Crystals,M.P. 163-164 C N=4.65 (4.46).

H CCH2CH2N(CH3)2-HCl S. 291 B-dirnethylaminoethyl-3-nitro-4-chlorophenylketonc hydrobromide Crystals, M.P. 183-184 C N=8.23(8.29).

S. 300 fl-morph01inoethyl-2-hydroxyphenylketonc hydrochloride Crystals,M.P. 194-195 C N= 5.27(5.15),

S. 366 fl-pipcridinoethyl-2-hydroxyphenyl ketone(B-piperidylcthyl-Z-hydroxy- Oil phenyl ketone).

S. 367 B-dimethylaminoethyl-4-chloro-1-naphthylketone- Oil S. 368B-dimethylaminoethyl-2-hydroxyphenyl ketonei1 S. 369B-morpholinoethyl-4-nitrophenyl ketonc hydrochloride Crystals, M.P.192-194 0 N =9.53(9.31).

S. 381 B-morpho1inocthyl-2-nitrophenyl ketone hydrochloride Crystals,M.P. 276-280 C N=9.47(9.31).

S. 382 fl-dimethylaminoethyl-Z-nitrophenyl kctone hydrochlorideCrystals, M.P. 263-265 C N=11.06(10.82).

s. 333 fl-morpholin0ethy1-4-bromo-1-naphthyl ketone hydrochloride.Crystals, M.P. 172-174" C N=4.12(3.64)

S. 385 B-morpholinoethyl-2-chlorophenyl ketone hydrochloride Crystals,M.P. 165 C N =4.98(4.82).

S. 386 fi-dimethylaminoethyl-Z-chlorophenyl ketone hydrochlorideCrystals, M.P. 164-164. 5 C N=6. (5.64).

S. 387 fl-morpholinoethylA-fluoro-l-naphthyl ketone hydrochlorideCrystals, M.P. 188189 0 N =4. 51(4. 32).

S. 388 B-morpholinoethy1-4-chloro-1-naphthylketone sulfate Crystals,M.P. loo-161 C Cl=8. 64(8.82).

I H SO =7.47(7.98).

S. 389 B-dimethylaminoethyl-4-fiuoro-1-naphthyl kctone hydrochlorideCrystals, M.P. 173-174 C N=5. 4201.97).

I Cl=12.59(12. 8).

S. 390 fl-morpholinocthyl 4-methyl-1-naphthyl ketone hydrochlorideCrystals, M.P. 176-177 C N=4. 47(4.3s).

. Cl=l1. 3301.08).

3, 414 fl-morpholinoethyl-9-anthranyl ketone hydrochloride Crystals,M.P. -166 C N=3.97(3.93).

TABLE IContinued Mark Compound Physical characteristics Analytical data,

percent S. 558 B-dimethylaminoethyl-Q-anthranylketone picrate. CrystalsS. 559 fl-dimethylaminoethyl-l-naphthyl ketone picr e Cry S. 560B-morpholinoethy1-4-ohloro-l-naphthyl ketone picrate Cry S. 564fl-dimethylaminoethyl-4-chl0ro-1-naphthyl ketone picrate Crystals S. 565B-dipropylaminoethyl-l-naphthyl ketone picrate Crystals S. 575fi-1norph0linocthyl-4-chloro-1-naphthy1ketoneferrocy Crystals V S. 576fimorpholinoethyl-l-naphthyl ketone picrate Crystals Zineb, WhiCh 1sused for comparison as a presently 25 TABLE 111 available fungicide, iszinc ethylenebis (dithiocarbarnic acid).

(l) ENDOTHERAPEUTIC AND IMMUNIZING ACTIVITY I Vine peronaspora(Plasmopara viticola) The field tests, which were carried'out in orderto confirm the preliminary results obtained on vines grown in aconditioned room, utilized the following method. In an open field, thelower face of vine leaves is sprayed with a conidia suspension insterile water, having a density of about 500,000 conidia per milliliter.After infection, the leaves are closed in polyethylene bags previouslywetted on the inside, in order to have a moisture-saturated atmosphere.'About 16 hours after the infection, the bags are removed and at givenintervals of time from the infection the leaves are subjected tospraying of both their upper and lower faces with an aqueous solution ofthe products under consideration.

After a period of time which varies from 4 to days after the infectionand depends on the temperature and moisture conditions, the leaves arecut and incubated in a room conditioned at C. and saturated withmoisture until the appearance of the fungus. The results of a first testare reported in Table II.

TABLE II [Endotherapeutical activity of '3-fl -aminoarylketones onPlasmopam m'tiaola. Treatment with a 5% dosage. -Results determined on10 leaves per thesis] '3=growth on one-half of the leaf surface 4 growthon two-thirds of .the leaf surface fizgrowth on the whole leaf surfaceThe influence of time elapsing between the infection and the treatmenton the endotherapeutical activity of the product was then studied. As asevere control, a particularly high inoculum density (800,000 conidiaper cc.) was purposely used so that, in the case of treatment effected 7days after the infection, the inner development of the fungus myceliumWas so considerable as to cause expanded necroses on wide areas of theleaf. The results were positive, as shown in Table III.

[Influence of time between infection and treatment on theendotherapeutic activity of 3-fl-aminocthylaryl ketone hydrochlorides.Treatment with 5% dosage. Infection in open field on vine leaves withPlasmoparaniticola] Fungus growth (see below) Interval of time betweeninfection and treatment: 7 days (average of 10 repetitions per eachthesis) Interval of time between infection and treatment: 3 days(average of 6 repetitions per each thesis) Product 0=no growth.

l=sporadic growth spots on the leaf lamina. 2=growth on of the leafsurface.

3= growth on of the leaf surface. 4=growth on of the leaf surface.5=growth on the whole leaf surface.

Moreover, the results of the field tests confirmed the results obtainedin the laboratory in the screening stage.

These results have shown that the antiperonospora activity of,G-aminoethylnaphthyl ketone hydrochlorides is markedly higher than thatof fl-aminoethylphenyl ketone hydrochlorides. As a substancerepresentative of the first class was chosen ,8 dimethylaminoethyl 1--naphthyl ketone (S. 210) and for the other classB-dimethylaminoethyl-Z- hydroxyphenyl ketone hydrochloride (S. 92), andp1- rnorpholinoethyl-Z-hydroxyphenyl ketone hydrochloride (S. 300). A 2%dosage of S. 210 resulted in a complete curative action within 3 daysafter infection with Plasmopara vitico la. Neither S. 300 or S. 92showed any curative action under the same conditions of infection andtreatment and after the same lapse of time.

A field trial was carried out with one of the more active products (S.236) to determine the relationship between the dosage and theendotherapeutic action. The infection was effected by spraying theunderside of the leaves with a natural inoculurn; the treatment with thevarious doses was carried out after 3 days. The results are reported inTable IV.

TABLE IV [Open field endotherapeutic activity of S. 236, applied invarious doses on .Plasmopara viticola; a natural inoculum was used forcarrying out the infection. Results determined on 5 leaves for eachthesis] Pathogen growth (see below) Product 0=no growth.

1 =growth traces.

2=sporadic traces of growth on the leaf face. 3=growth on of the leafsurface. 5=growth on of the leaf surface.

Apple scab (Venturia inaequalisf.c. Fusicladium dendriticum) TABLE VOpen-field endotherapcutic activity of some products of the series otB-aminoethylaryl ketones on apple trees previously infected with conidiaof Fusicladium dendriticum (pathogen of apple scab)] Ooricen- Percentageof damaged leaf tration, surface in comparison with percent controlconsidered as Product rowccwwwmcb 9999K??? omocaocmq French bean rust(Uromyces appendicolatus) French bean leaves, cut at the base of thestem, are incubated in Petri dishes having a diameter of 20 cc.,containing 2 filter paper discs (separated by 6 glass rings). The leaf,with its upper face upward, is placed on the upper disc. The upper discis provided With a central hole. A drop of a test product solution ofknown volume and concentration is placed on the leaf by means of amicrosyringe-in correspondence With the hole in the upper disc. Afterdeposition of the drops, the dishes are kept open to allow a perfectdrying of the deposit. The dishes are then kept closed for hours, toallow the penetration and diffusion of the product. The dishes are thenopened, and the leaves are upset with the lower face upward, so that theapplication point of the drop corresponds to the center of the hole inthe disc. Thereupo-n the leaves are infected by spraying on theirunderside a suspension of U. appendiculatus spores in sterile water,with a density of about 150,000 spores per cc. After infection, thedishes are opened and the contents incubated in a room conditioned at C.for a period of 7 to 10 days to permit the complete growth of thefungus. The activity of the product is evaluated on the basis of thesize of the zone of inhibition of the fungus growth, in correspondencewith the point of application of the drop. The results obtained by thesaid technique are reported in Table VI.

TAB LE VI Immunizing activity of a series of fl-aminoethylaryl ketonesapplied as drops onto French bean leaves infected with Uromycesappendiculatus] Activity Activity Product (inhibition Product(inhibition halo in cm.) halo in cm.)

S. 5-6 S. 127 1-1. 5 S. 4 S. 1-1. 5 S. 4 S. 1-1. 5 S. 4 S. 1-l.5 S. 4 S.1-1. 5 S. 4 S. l-l. 5 S. 4 S. 1-1. 5 S. 4 S. l-l. 5 S. 4 S. l-l. 5 S. 4S. 1-1. 5 S. 4 S. l-l. 5 S. 2-3 S. 1-1. 5 S.- 2-3 S. 1-1. 5 S. 2-3 S.1-1. 5 S. 2-3 S. 1-1. 5 S. 2-3 S. l-l. 5 S. 2-3 S. 1-1. 5 S. 2-3 S. 1-1.5 S. 2-3 S. l-l. 5 S. 2-3 S. 1-1. 5 S. 2-3 S. 1-1. 5 S. 2-3 S. 1-1. 5 S.2-3 S. 1-1. 5 S. 2-3 S. 1-1. 5 S. 2-3 S. l-l. 5 S. 2-3 S. 1-1. 5 s. 2-3s. 1 -1. 5 S. 2-3 S. 1-1. 5 S. 2-3 S. 1-1. 5 S. 2-3 S. 1-1. 5 S. 2-3 S.l-l. 5 S. 2-3 S. 1-l. 5 S. 2-3 S. 1-1. 5 S. 1-1. 5 S. 1. 1. 5 S. 1-l. 5S. 1-1. 5 S. 1-1. 5 S. 1-1. 5 S. 1-1. 5 S. 1-1. 5

Tests were also carried out in order to evaluate the systemic immunizingactivity on French bean plants infected with U. appendicula'tus. 6-cm.high French bean plants, growth in a pot in a conditioned room, aretreated by spreading -a paste formulati-on containing 3% (Table VI I)and 5% (Table VIII) of active ingredient onto the stem zone comprisedbetween the base and the insertion point of the cotyledons. Thecomparative evaluation of the migration of the products is carried outby taking and infecting with U. appendiculatus all leaves at variousintervals from the treatment. The results are determined by counting theuredospores in each leaf. In Tables VII and VIII are summarized theresults obtained by operating with some ,B-aminoethylaryl ket-ones onFrench bean plants.

TABLE VII Disease percentage Interval of time between the referred tothe control treatment and the infection, days Tested leaves S S. S. 92210 300 12. 9 0. 0 0. 0 0. 0 0. 0 0. 0 Tertiary 0. 0 0. 0 O. 0

(The control was 100.)

In stemrapplication tests,

containing of active ingredient have previously been applied. Intervalof time between the treatment and the infection 7 days] Diseasepercentage referred to the control Product Primary Secondary Tertiaryleaves leaves leaves Analogous results were obtained in field tests.Under these conditions the products are brushed over an area of 30 cm.on the base portion of the stem of 1.70 m.-high French bean plants. Fourdays after the treatment (carried out with a paste compositioncontaining 10% of active ingredient), the plants3 repetitions pertestare infected with an aqueous suspension of U. appendiculatusuredospores (180,000 per 00.). The plants are then covered by apolyethylene bag for 16 hours, in order to favor the growth of thepathogen. The results are determined by counting the uredosporesdeveloped on'each leaf. The results obtained are reported in Table IX.

- TABLE IX [Immunizing activity obtained in field tests against Uromycesappendicitlatus on French bean plants spread on the stem with 2fl-aminoethylarylketone hydrocliloridesl Disease percentage Productreferred to the control Tests for evaluating the systemic activity byroot absorptionare carried out by immersing the roots of -dayold Frenchbean plants, carefully washed with distilled Water, in glass vesselscontaining 200 cc. of an aqueous solution of the products to be tested.After 44 hours of immersion the roots are carefully washed and thesolution is replaced by distilled water. The evaluation of theabsorption, migration and activity is carried out by removing the leavesat various intervals of time after treatment and placing them in a wetchamber Where they are infected with U. appendiculatus by the usualprocess, then determining the disease index in respect to the control.The results are reported in Table X.

TABLE X [Immunizing activity against Uromyces appendz'culatus ofZfi-aminoethylphenyl ketone hydrochlorides applied by root absorption(solutlion of 200 ppm.) to French bean plants grown in a conditionedroom Disease percentage Product referred to the control (6 leaves perthesis) S. 92 0.2 S. 0. 3 Control 100. 0

Root absorption tests were also carried out in open field on 1.7-rn.high French bean plants (cv. Borlotto) in groups of 3 plants. Thetreatment is carried out by spraying the soil in a circular zone of20-30 cm. around the stems of each group of 3 plants with an aqueoussolution containing 3% of active product. Three treatments are effected,at one day intervals, by administering 5 liters of liquid per plantgroup and per treatment. Two days after the last application, the lowerface of cut leaves were sprayed with U. appendiculwtus uredospores(200,000 per cc.). The leaves are then incubated in Petri dishes at 20C. until the uredospores are formed. The results are determined bycounting the uredospores formed in each leaf. These results are reportedin Table XI.

p TABLE XI [Immunizing activity results obtained with S. 210 on Uromycesappendiculatus, by applying the product through root absorption ontoFrench bean plants in open field] Disease percent referred to thecontrol T1 sated plants 3 0. L5 Control plants. 352 100 Experimentalthesis Carnation rust (Uromyces dianthi) 20-cm. high carnation plants,grown in a conditioned room and :artifically infected with a suspensionof Uromyces dianthi spores (density 200,000 spores per 00.). Theinoculated plants are kept for 48 hours in a moisturesaturated room at20 C. and are then subjected to the treatment with the products in a 1%aqueous solution. The plants are then incubated at 20-25 C. with 50 to70% of relative humidity till the fungus growth. The determination ofthe results (18 days after the infection) is carried out by counting theuredospores formed on each leaf. These results are reported in TableXII.

TABLE XII [Endotherapeutic activity of a series of products belonging tothe class of fl-aninloethylaryl ketones against Uromyces dianthi] Disease percent referred to the I Number of repetitions per test Averagepustule number per plant Plant Product Alba Bianco Venere.

' NINHOM PE".

Olive anthracnose (Gloeosporium olivarum) To determine the immunizingactivity of the products to be tested in respect of Gloeosporiumolivarum, the following procedure was adopted. Olives (cv. Leccino) aresprayed at low volume with a =microsprayer operating at air pressure of1.6 atrn. By spraying for 30 seconds, a solution of the test fungicidewith a concentration of 5% in distilled water, each drupe is coveredwith a deposit of 30 of active substance. -T he olives thus treated arekept at room temperature for 4 days. Thereafter the drupes are washedwith distilled water 3-4 times (in this way it has been ascertainedexperimentally that all the residue of the active substance present onthe drupe surface is removed) and the pathogen is then inoculated asfollows:

(1) Suspensions in sterile distilled water are prepared 'with G.olivarum conidia obtained from a 4-day culture developed on potatodextrose agar Dipco at 21 C.; the density of the suspension is broughtto a value of 700,000 conidia/ cc.;

(2) Each drupe is wounded in 5 points (in the apex and in otherdiametrically opposed four points on the drupe surface) by passing theolive on a very fine abrasive paper so as to cause a lesion involving acircular zone having a diameter of about 2 mm. and a depth of amillimeter fraction;

(3) On each lesion made as described above (2), a fine drop of conidiasuspension prepared as above (1) is deposited.

After inoculation, the olives are incubated for 2 days in a wet room at26 C. and then for 4-5 days at 21 C.

TABLE XIII [Immunizing activity of some products on pick-up olivesinoculated with Gloeosporium olivarum, 4 days after the treatment]Disease percentage referred to the control Product Concentration (2)PREVENTIVE SPORICIDAL ACTIVITY BY CONTACT OR FUMIGATION Down mildew ofgrape (Plasmopara viticola) Tests for determining the preventivesporicidal activity by contact were carried out on vine plants grown ina conditioned room by operating as follows. The products, in aqueoussuspension or solution, are sprayed on the lower face of the leaves ofhomogeneous groups of plants. 24 hours after the treatment, the plantsare infected by spraying onto the lower face of the leaves a suspensionof Plasmopara viticola conidia (400,000 per cc.) and then incubated for24 hours at 20 C. in a room saturated with humidity. The plants aretransferred to a room at 2025 C. Two days later the group of test plantsis transferred into a room saturated with humidity until the fungus iscompletely grown. The results are taken by determining on each leaf thepercentage of surface injured by the pathogen. In Tables XIV, XV, XVIand XVII are reported the results obtained in successive tests.

TABLE XIV [Preventive sporicidal activity by contact of someB-aminoethylaryl ketones against P. viticola (3 repetitions for eachtest)] Percentage of leaf surface injured by the pathogen referred tothe control=100 Product 111 p.p.m. 36 p.p.m. 12 p.p.m.

rue- TABLE XV [Preventive sporicidal activity by contact of S. 300 andZineb agains P. viticola (3 repetitions per test)] [Preventivesporicidal activity by contact of S. 236, S. 205 and Zineb against P.viticola (3 repetitions per test)] Percentage of leaf surface injured byEhe pathogen referred to the con- Product 0l=100 2O p.p.m. 10 p.p.m. 5p.p.m.

TABLE XVII [Preventive sporicidal activity of S. 558, S. 560, S. 564, S.565 and Zineb against P. viticola (3 repetitions per test)] Percentageof the surface of the leaf covered with Product the mold NQH Zineb Thepreventive sporicidal activity by fumigation against the same pathogenwas determined by the following method. Two superposed filter paperdiscs having a diameter of 23.5 cm. and provided with a central holewith a diameter of 8 cm. are placed on a perfectly flat glass sheet andthen impregnated with an aqueous suspension or solution of the productto be tested. Immediately thereafter, in correspondence with the centralhole, is placed a pot containing a 25-cm. high vine plant whose leaveshave previously been infected with Plasmopara viticola (the infectiontechnique is the same as described above). The two operations (treatmentof the paper discs and inoculation) are carried out by two personsoperating simultaneously so as to allow the almost immediate covering ofthe inoculated plant and of the two underlying discs with a glass cuphaving a diameter of 28.5 cm. and a height of 44 cm. The plantsprotected by the cups are incubated. at 20 C. for 24 hours, thenuncovered and transferred at 2025 C. for further 2-3 days and finallykept at 20 C. in a room saturated with humidity till the pathogen isgrown.

The determination of the results is carried out by evaluation of theleaf surface covered by the fungus. In Table XVIII are reported thevalues determined by the said method.

TABLE XVIII [Preventive sporicidal activity by fumigation exerted invivo by some fi-aminoethylaryl ketones against P. viticola (1.9 mg. ofactive substance applied per 1000 cc. of room volume), one vine plantper each test] Percentage of leaf surface injured by the pathogen,

referred to the controls= (average of 2 tests) Product oocaccc After thetest, phytotoxic effects were observed only in the case of S. 300.

I Apple scab (Fusicladium dendriticum) TABLE XIX [Preventive sporicidalactivity by contact of S. 280 and Zineb against Fusicladium pirim m as afunction of the use doses and the interval of time elapsing between thetreatment and the infection (two plants for each test)] Percentage ofleaf surface injured by the pathogen. referred to the contrls=100Interval of time between treatment and infection, days Product Bean rust(Urornyces appendiculatus) The evaluation of the sp-oricidal activity bycontact was carried out by adopting the following technique. Thetreatment is effected on l415-day-old plants grown in a pot in aconditioned room. The test products are sprayed in aqueous solutionswith various concentrations onto both faces of the primary leaves. After24 hours, the plants are inoculated with a suspension of uredospores(180,000 spores per cc.) and incubated for 48 hours in a room saturatedwith moisture at C.; the plants are then transferred into a room at20-25" C. with a relative humidity of 5070% until the pathogen is grown.

The results were taken by counting the number of blisters developed oneach leaf. The values determined in two tests are reported in Tables XXand XXI.

[Preventive sporicidal activity by contact exerted by S. 210 and S. 300

against Uromyces appendiculams] Disease index r3ferred to thecontro1s=100 plants per test) Product 100 ppm; 50 p.p.m. 25 p.p.rn. 12.5p.p.m.

By using a technique similar to that already described for the -in vivotest with the Plasmopara viticola, the preventive sp'oricidal activityby fumigation was evaluated also on French bean plants infected withUromyces dpperidiculdtus. Q

is In Table XXII are reported the values obtained with variousB-aminoethylaryl ketones.

' TABLE XXII [Preventive sporicidal activity by fumigation exerted invivo by a series of fi-aminoethylaryl ketones against U. appen liculatus(2.9 mg. of 1actgrle substance applied per 1000 cc. of room volume) (oneplant per Product Disease index referred to the controls ocoo opoopooooooccoooomcpco op HOOP r- In Table XXIII are values obtained ofthe preventative sporicidal fungicide activity of several of the,B-aminoethylaryl ketones.

TAB LE XXIII [Preventative sporicidal activity of S. 558. S. 559 andZineb against Ummyces appmiiculatus on beans (16 repetitions perdosage)] From the values reportedin Tables XXII and XXIII it is evidentthat the tested products more or less inhibited the development of thebean rust. This particular type of' activity is to be ascribed to adirect action against the pathogen and not as an influence on thereceptivity of the plant, as it was demonstrated (for some compounds ofgreater interest) by operating on U. ap-

pendiculalus spores placed as drops onto slides to let them germinateunder conditions similar to those described a-bove in the case of thetests carried out by fumigation in vivo. 1

Intwo tests carried out at different times the results reported in TableXXIV were obtained. I I i I I TABLE XXIV [Preventive sporicidal activityby iurnigationexerted in vitro by S. 92, S. 192, S. 210, S. 212, S. 236and S. 300 against Uromyccs appem diculatus uredospores (10 mg. ofactive substance administered per 1000 cc. of room volume)] Carnationrust (Uromyces dianthi) The test for evaluating the preventivesporicidal activity by contact was carried out on carnation leaves, cutat their base, by operating as follows. Both the upper and lower facesof the leaves were sprayed with aqueous solutions or suspensions of thetest products. The leaves supported by a net, are placed in'boxescontaining a layer of distilled water on the bottom, so that the base ofeach leaf remains immersed to a depth of about It cm.

As soon as the sprayed deposits are dried, the leaves are inoculated byspraying with an uredospores suspension (200,000uredospores per cc.).The boxes are then covered and the contents incubated at 20 C. until thefungus is grown, and the number of uredospores on each leaf is counted.In Table XXV are summarized the results obtained by operating as above.

TABLE XXV Disease index referred to the control (considered as 100) av-Product erage of 10 repetitions l p.p.m. p.p.m.

S. 3. 80 G. 60 S. 1. 90 6. 60 S. 1. 98 7. 92 S. 0.57 1. 70 S. 2.80 3.80s, 0. 60 1. 30 S. 1.90 2. 60 S. 1. 99 4. 95 S. 0 1. 98 S. 0 1. 99 S.1.14 3. 99 S. 0. 60 2. 60 S. 2. 80 18.00 S. 1. 98 4. 95 S. a 3.50 5.30S. 1. 2. S. 1. 3. S. 0.70 2. S. 1. 40 3. 30 S. 1. 10 4. 00 Z 2. 50 10.00

The fumigation activity of some of the compounds was also determined inrespect to Uromyces dianthi by operating on single carnation leavesimmersed with their base in a vial containing distilled water andincubated in a vessel having a volume of 1800 cc, The bottom ofthe'ves-sel contained (at the distance of 2-3 cm. from the vials) afilter paper disc with a diameter of 18 cm., impregnated with a 1.5%aqueous solution of the test prodnot (10 cc.). The leaves were infectedjust before the treatment. The closed vessel is then incubated at 20 C.for 48 hours and then opened at 20-25" C. Under these conditions (8.33mg. of active substance administered per 1000 cc. of room volume), thefollowing disease percent indexes were obtained:

S. 210S. 300S. 192:0 S. 212--S. 236==0.4 Controls: 100

Blue mold (Peronospora tabacina Adam) Leaves cut from hydroponic tobaccoplants have been used for testing the surface protective activityagainst P. talyacina. The petioles of leaves have been soaked during theentire testing period in a nutritive solution. The contamination of theleaves treated with the compounds to be tested has been achieved byspraying a conidial suspension (about 100,000/cc.) on the upper side ofthe leaves. The infected leaves have been transferred 'into a glass bellat 100% humidity and 15 C. Twentyafour hours later the leaves have beentaken out from the bell in order to let them dry and then they have beenkept at 20 C. and 1,00% humidity until fructifications have raised. Theresults have been checked 8 days after the infection by the evaluationof the percentage of the leaf surface covered with the mold.

The results of the above test are given in Table XXVI.

TAB LE XXVI [Protective surface activity of S. 558, S. 559, S. 560, S.564, S. 565, S. 575, S. 576 and Zineb against P. tabaeina on detachedleaves of tobacco (4 repetitions per dosage)] Fungus growth (see below)Concentration of active ingredient, p.p.m.

Product Lower surface of Upper surface of the leaf the leafWOOHOHOOONHOOCOOOHOOOHOOONl-OOHOQO 0 =no growth.

1 =sporadic growth.

2=growth on M of the leaf surface. 3=growth on of the leaf surface.4=growth on of the leaf surface. 5=growth on the entire surface.

Phyt0t0xicity.-As concerns the phytotoxicity of the instantparasiticidal compounds (under phytotoxic is meant the complex ofmorphological and-physiological changes deriving from the application ofthe products to the plants), it has been found that most of the testedproducts do not have any appreciable phytotoxic effect when applied inthe concentrations which are useful for a full entdotherapeutic andimmunizing action, inside the vegetal tissues.

However, we can not neglect considering the phytotoxic action, if any,occurring from the residues which could possibly remain outside theplant, inasmuch as the,

phytotoxic activity of the residues could also be modified by reactionscaused by atmospheric conditions.

The phytotoxicity of the instant products was therefore evaluatedfromthe results of tests carried out purposely in open field and fromcareful observations during field tests carried out to determine thepreventive and endotherapeutic and immunizing activities of theseproducts.

The phytotoxicity tests for determining the non-phytotoxic limit dose onsome products according to the-invention were carried out on flowers andleaves of some cultivated varieties of pears and apples. The productsdissolved in distilled water were sprayed over small branches of plantsgrown in open field, and the phytotoxic effect was evaluated 8 daysafter the treatment.

Five tests were carried-out in May and June, under similar weatherconditions in both months. The results are reported in Table XXVII.

TABLE XXVII From the various tests it appeared that all the productsexamined did not show any specific toxicity even at the highest dosetested (500 mg./ kg. of living animal).

We claim:

1. The process of applying beta-dimethyl-aminoethyl- 9-anthranyl ketonepicrate to a plant, whereby said plant is immunized to fungus growth.

2. A process of immunizing a plant against fungus growth which comprisesapplying a beta-dimethyl-aminoethyl-l-naphthyl ketone picrate to saidplant.

3. A process of immunizing a plant against fungus growth which comprisesapplying beta-morpholino-ethyl- 4-chloro-1-naphthyl ketone picrate tosaid plant.

4. A process of immunizing a plant against fungus growth which comprisesapplying a beta-dimethyl-aminoethyl-4-chloro-l-naphthyl ketone picrateto said plant.

5. A process of immunizing a plant against fungus [Non-phytotoxic limitdose of a. series of B-aminoethylaryl ketones applied in open field onleaves of some cultivated pear and apple varieties] Non-phytotoxic limitdose Date of Pears Apples Product treatment Butirra Decana Passa GoldenRosa Ranetta o1 clairgeau dinverno crasdelicious Manto- Canada sana vanaS. 4. 5 1. 25 1. 25 1. 25 0. 62 2. 50 2. 50 S. 4. 5 0. 62 1.25 1.25 0.625.00 10.00 S. 4. 5 1. 25 1. 25 1. 25 0.62 10.00 10.00 B 12. 5 0. 62 2.50 5. 2. 50 5. 00 5. 00 S. 12.5 1. 25 1. 25 5.00 5. 00 10.00 5.00 S.12. 1. 1. 25 10.00 5.00 10.00 10.00 S. 26. 5 1. 25 1. 25 1. 25 5. 0010.00 10. 00 S. 26. 5 1. 25 0.62 1. 25 5.00 5.00 5.00 S. 26. 5 1. 250.62 1.25 1. 25 1. 25 10.00 S. 4. 6 1. 25 1. 25 5.00 10. 00 10.00 10.00S 4. 6 1. 25 1. 25 5.00 10.00 10. 00 10. 00 S. 4. 6 1. 25 1. 25 5.0010.00 10.00 10.00 S 25. 6 10. 0O 5. 00 10. 00 10. 00 10. 00 10. 00 S25.6 1. 25 2. 2. 50 2. 50 10.00 10.00

When dosages were applied that were higher than the highestnon-phytotoxic dosages, circular colored zones only were noted. Thesezones were more frequent on the lower than on the upper side of theleaves. Moreover, sprayings carried out with 2% aqueous solutions of S.210 and S. 212 onto apple trees in bloom (cv. Abbondanza) grown on anespalier, caused marginal necrosis on portions of the petals withoutimpairing the thriving. Also phytotoxicity tests carried out on oliveleaves and fruits with the various products of the invention in aqueoussolutions containing 5% of active substance, did not show any phytotoxicactivity 5 days after the treatment.

The phytotoxicity tests on vines of S. 210 and S. 212, which underlaboratory conditions have shown the highest antiperonospora activity,were carried out in open field in order to determine the phytotoxicactivity, if any, on grapes in different growth stages, with aqueoussolutions containing 2% of active substances. The applications were madeon vines -of the cv. Sangiovese before blooming and immediately afterthe beginning of grape formation. From repeated observations of thetreated vines no phytotoxic effect caused by the tested products wasnoted.

(3) TOXICITY ON WARM-BLOODED ANIMALS Tests were carried out to determinethe amount of the above products to produce acute toxicity, if any, byingestion by warm-blooded animals. The research was limited to somecompounds (S. 169, S. 210, S. 212 and S. 236), using both sexes of thewhite albino mouse as test animals. Products S. 210 and S. 21 2 weresufficiently water-soluble to be administered as solutions in distilledwater; S. 169 was administered in a 50-50 solution of water andmethylacetamide; S. 236 was admin istered in aqueous suspension.

growth which comprises applying beta-dipropyl-aminoethyl-l-naphthylketone picrate to said plant.

6. A process of immunizing a plant against fungus growth which comprisesapplying beta-morpholino-ethy1- in which A is aryl selected from thegroup consisting of simple and substituted phenyl, naphthyl, andanthranyl,

wherein the substituents are taken from the group consisting ofhydroxyl, halo, nitro, alkyl and oxyalkyl; Y is selected from the groupconsisting of morpholino, piperinino and in which R and R are selectedfrom the group consisting of hydrogen and lower alkyl to systemicallyimmunize the plant'against fungus growth.

(References on following page) References Cited by the Examiner UNITEDSTATES PATENTS Treboux 167-22 Horst 167-33 D Swaine 167-22 Epstein et a1167-22 Schultz 260-5705 Moed et a1 260-5705 Gregory 167 30 D Eden 167-306/ 1960 Darlington 167 30 10/1960 Hopkins et a1 167-30 OTHER REFERENCESI. Am. Chem. Soc., 64 (1942), 453. J. Org. Chem, 10 (1945), 259.

J. Org. Chem., 10 (1946), 215.

Org. Sytheses, 1946, vol. 23, page 30.

8. A PROCESS OF IMMUNIZING A PLANT AGAINST FUNGUS GROWTH WHICH COMPRISESAPPLYING TO A PLANT A PICRATE OF FERROCYANIDE OF A COMPOUND OF THEFORMULA